Apparatus and method for the separation and/or formation of immicible liquid streams

ABSTRACT

Apparatus and method are provided for the substantial separation on a continuous flow basis of immisicible liquids flowing as a common stream, with one of said liquids substantially encapsulating another of said liquids, in a flow conduit. The conduit comprises an inner surface which is selectively &#34;wettable&#34; by one of the liquids to the substantial exclusion of another of the liquids; and the liquid separator apparatus are disposed in the conduit and operate to substantially separate the liquids, or at least a portion of one of the liquids, from the common liquids stream as that stream flows into contact with the separator apparatus in the conduit. 
     Apparatus and method for the introduction of a liquid on a continuous flow basis into a flowing stream of another liquid which is immisicible therewith and is flowing in a flow conduit are also provided. The conduit again comprises an inner surface which is selectively &#34;wettable&#34; by one of the liquids to the substantial exclusion of the other of the liquids; and the liquid introduction apparatus are disposed in the conduit and operate to form a common stream of the liquids for continued flow in the conduit, with one of the liquids being substantially encapsulated by another of the liquids, as the liquids are merged at the liquid introduction apparatus in the conduit. 
     A liquid processing system including the conduit is provided wherein the liquid introduction apparatus are utilized to form the common liquids stream with one of the liquids being substantially encapsulated within the other of the liquids at a first location in the system; and the liquid separator apparatus are utilized to substantially separate the liquids, or at least a portion of one of the liquids, from the common liquids stream at a second location in the system which is downstream of the first location. 
     As representatively configured in the herein disclosed preferred embodiments, the apparatus and method are applied to automated continuous flow sample liquid analysis systems wherein one of the liquids is an aqueous sample liquid, and the other of the liquids is an isolation liquid which selectively &#34;wets&#34; the hydrophobic inner surface of the flow conduit to the substantial exclusion of the aqueous sample liquids, and which functions to substantially encapsulate the aqueous sample liquids in the common liquids stream; and, for such application, the apparatus and method provide for significant increase in the accuracy of the sample liquid analysis results.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to new and improved apparatus and method for thesubstantial separation and/or formation of immiscible liquid streams ona continuous flow basis; and which are particularly adapted to use inautomated, continuous flow sample liquid analysis systems to maximizethe accuracy of the sample liquid analysis results.

2. Description of the prior art

Although a number of methods and apparatus are known for the separationof sample liquids from immiscible isolation liquids within which thesame are encapsulated for minimization of sample liquid carryoverattendant sample liquid analysis, these will generally be found to relyprimarily upon the natural separational effects of the differences inspecific gravity between those liquids; and thus will not be foundeffective to accomplish the virtually immediate, and complete in termsof totally distinct locations of the thusly separated liquids,substantial separation of the liquids on a continuous flow basis asinherently provided by the apparatus and method of this invention.

More specifically, U.S. Pat. No. 4,121,466 issued Oct. 24, 1978 to AllenReickler, et al for "Liquid Dispenser With An Improved Probe" andassigned to the assignee hereof, discloses the use of an immisciblehydrophobic isolation liquid to encapsulate successive aqueous sampleliquids for minimization of sample liquid carryover attendant sampleliquid analysis. In this apparatus, wherein the isolation liquid is ofgreater density than the sample liquids, the former is simply allowed tosettle out from the latter to the bottom of a reaction receptacle intowhich the isolation liquid-encapsulated sample liquids are dispensed,thereby leaving the sample liquid readily accessible for reaction withreagent liquids as may then be introduced into the receptacle. Thesettling out of the isolation liquid can and does take time and, in anyevent, leaves the thusly separated isolation and sample liquids inessentially the same location, e.g. the reaction receptacle.

In like manner, U.S. Pat. No. 4,357,301 issued Nov. 2, 1982 to MichaelM. Cassaday, et al for "Reaction Cuvette" and assigned to the assigneehereof, also discloses the use of an immiscible isolation liquid toencapsulate successive aqueous sample liquids for minimization of sampleliquid carryover attendant sample liquid analysis. In this apparatuswherein the isolation liquid is again hydrophobic and apparently ofgreater density than the sample liquids, sharp projections or the likeof a hydrophilic material are provided at the bottom of the reactioncuvette, and operate to puncture the isolation liquid-encapsulatedsample liquids as the same are introduced into the cuvette; therebyfreeing for reaction the sample liquids from the isolation liquid whichsimply floats to the top of the cuvette. Again, this separation can anddoes take time, and in any event, leaves the isolation and sampleliquids in essentially the same location, e.g. the reaction cuvette.

Under the circumstances, it has been determined by applicants that thecontinued presence of the "separated" isolation liquid with the sampleliquid at the same location can and does present significant problemswith regard to the accuracy of subsequent sample liquid analysisresults; and especially in those instances wherein those sample liquidanalysis results are arrived at through use of sample liquid analysismethodologies involving, for example, reflectance spectroscopy, ionselective electrodes, colorimetry, cell counting and or enzyme coiloperation.

Hydrophobic filtration, for example as disclosed by the "nonwet" filterin U.S. Pat. No. 4,266,559 issued May 12, 1981 for David S. Akhavi for"Blood Sampler," wherein a filter of hydrophobic material is used toprevent the escape of an aqueous sample liquid from a collection devicewhile permitting the passage of air therethrough to enable filling ofthe device, is also known in the prior art, but is clearly totallyirrelevant to the substantial separation of immiscible liquids from aflowing stream to distinct locations on a continuous flow basis.

Also of limited relevance to liquid separation are conventionaldebubbler devices as have now become standard in continuous flow sampleliquid analysis systems and which operate to remove the air segmentsfrom a continuously flowing, air segmented sample liquid stream prior tosample liquid analysis. These debubbler devices, which operate primarilyon the very significant differences in specific gravity between air andthe sample liquids of interest are clearly totally inapplicable to theeffective substantial separation of immiscible liquids on a continuousflow basis.

With regard to immiscible liquid stream formation, it is known in theprior art to form immiscible liquid streams on a continuous flow basisby the essentially concomitant introduction of aqueous sample liquids,and a hydrophobic isolation liquid which is immiscible therewith into ahydrophobic flow conduit which is selectively "wettable" by theisolation liquid to the substantial exclusion of the aqueous sampleliquids, thereby essentially encapsulating the former in a layer of thelatter and effectively minimizing aqueous sample liquid carryover.

More specifically, in most instances, this will be seen to beaccomplished by the concomitant aspiration of the aqueous sample liquidsand the immiscible isolation liquid into the sample analysis system bythe sample aspirating probe as disclosed, for example, in U.S. Pat. No.4,121,466 as discussed hereinabove, and in each of the U.S. Pat. No.4,253,846 issued Mar. 3, 1981 to William J. Smythe, et al for "Methodand Apparatus for Automated Analysis of Fluid Samples," and assigned tothe assignee hereof, and U.S. Pat. No. 4,517,302 issued May 14, 1986 toSteven Saros, et al for "Sample Analysis System," and also assigned tothe assignee hereof. Although prior art apparatus of this nature dooperate to satisfactorily continuously form the isolation liquidlayer-encapsulated, sample liquid stream, the same are of course limitedin location to the aspirating probe at the entry point of the analysissystem without the system flow conduit, thereby limiting the versatilitythereof. In addition, since these prior art apparatus are strictly tiedinto the operation of the analysis system aspirating probe, satisfactoryoperation thereof by definition requires the use of moving parts whichcan prove somewhat problematical. Also, the extremely high speeds ofoperation of contemporary sample liquid analysis systems of the naturehere under discussion require extremely high speeds of aspiration probemovement through decidedly limited distances, and these requirements canadversely affect the satisfactory formation of the isolation liquidlayer by the probe.

U.S. Pat. No. 3,479,141 issued Nov. 18, 1969 to William J. Smythe, etal, and now expired, for "Method and Apparatus for Analysis," andassigned to the assignee hereof, discloses the formation on a continuousflow basis of an isolation liquid layer encapsulated sample liquidstream by the concomitant pumping through separate compressible pumptubes of a peristaltic pump of immiscible isolation and buffer liquidsto a tube junction for merger therein and flow therefrom through therecipient side of a dialyzer to acquire the sample liquids forsubsequent flow of the thusly isolation liquid encapsulated sampleliquid stream through a glass conduit to a hydrophobic conduit foradditional sample liquid processing. Formation of the isolationliquid-encapsulated sample liquid stream in this manner of necessityresults in an extremely "rich" isolation liquid layer, to verysignificant economic disadvantage as made clear by FIG. 1 of the drawingof that patent; and operates to effectively minimize sample liquidcarryover only upon the arrival of the stream at the hydrophobicconduit. In addition, formation of the isolation liquid-encapsulatedsample liquid stream in this manner is, of course, unduly complex, andsimply cannot in any event provide the precision of isolation liquidlayer formation as required by more contemporary, highly technicallysophisticated continuous flow sample liquid analysis systems.

U.S. Pat. No. 3,726,297 issued Apr. 10, 1973 to Richard H. Heimann,Aaron Kassel and Donald F. Kopelman for "Method And Device ForIntroducing For Mixing A First Liquid Into A Second Liquid," andassigned to the assignee hereof, discloses a tube disposed within anelongated passageway in which a first liquid is flowing for introducinga second liquid thereinto for mixture with said first liquid. In thisdevice, the first and second liquids are, of course, not immiscible; andthe second liquid is introduced into the first liquid in thecountercurrent direction relative to the flow of the latter.Accordingly, it will be immediately clear to those skilled in this artthat the device of U.S. Pat. No. 3,726,297 is totally inapplicable tothe formation of an isolation liquid-encapsulated sample liquid streamfrom immiscible isolation and sample liquids in a conduit which isselectively "wettable" by the isolation liquid to the substantialexclusion of the sample liquid.

OBJECTS OF THE INVENTION

It is, accordingly, an object of our invention to provide new andimproved apparatus and method for the substantial separation on acontinuous flow basis of immiscible liquids which are flowing in acommon stream in flow conduit means.

It is another object of our invention to provide new and improvedapparatus and method as above which are operable to separate thoseliquids to a very high degree of separational efficiency.

It is another object of our invention to provide apparatus and method asabove which are operable to effect the virtually immediate separation ofthe liquids from the flowing common liquids stream.

It is another object of our invention to provide apparatus and method asabove which are operable to separate said liquids, or at least asignificant portion of one of the liquids, to distinct and spacedlocations without contact therebetween.

It is another object of our invention to provide apparatus and methodoperable as above wherein one of said immiscible liquids issubstantially encapsulated within a layer of the other of said liquidsin said common flowing stream.

It is another object of our invention to provide apparatus and method asabove wherein one of said immiscible liquids is an aqueous liquid whichis to be separated from the other of said liquids.

It is another object of our invention to provide apparatus and method asabove which are particularly adapted to the separation of immiscibleliquids on a continuous flow basis in an automated sample liquidanalysis system wherein said conduit means are hydrophobic, one of saidimmiscible liquids is a sample liquid, and the other of said liquids isa hydrophobic isolation liquid which substantially encapsulates the samefor minimization of sample liquid carryover in said sample liquidanalysis system.

It is another object of our invention to provide apparatus and methodfor introducing one or more liquids on a continuous flow basis into aflowing stream of a liquid which is immiscible therewith to form acommon flowing stream of said immiscible liquids.

It is another object of our invention to provide liquid introductionapparatus and method as above wherein the thusly formed common stream ofsaid immiscible liquids will comprise a stream of one of said liquidssubstantially encapsulated within a layer of the other of said liquids.

It is another object of our invention to provide apparatus and method asabove which are operable to precisely form said encapsulating liquidlayer to a predetermined minimum thickness commensurate with thefunction thereof, thereby minimizing the amount of said encapsulatingliquid required for such function.

It is another object of our invention to provide liquid introductionapparatus and method as above which are operable to provide a wide rangeof predeterminable thicknesses for said encapsulating liquid layer, andwhich enable the formation of that encapsulating liquid layer to aprecisely predetermined, minimum effective thickness within that rangeto thereby minimize the required amount of said encapsulating liquid tosignificant economic advantage.

It is another object of our invention to provide liquid introductionapparatus and method as above wherein one of said immiscible liquids isan aqueous liquid which is to be substantially encapsulated in a layerof the other of said liquids.

It is another object of our invention to provide liquid introductionapparatus and method as above which are particularly adapted to theformation of an immiscible liquid stream on a continuous flow basis inhydrophobic flow conduit means of an automated sample liquid analysissystem wherein one of said immiscible liquids is an aqueous sampleliquid, and the other of said liquids is a hydrophobic isolation liquidwhich substantially encapsulates the same for minimization of sampleliquid carryover in said sample liquid analysis system.

It is another object of our invention to provide liquid introduction andliquid separation apparatus and method as above which are, in eachinstance, of particularly simple and straightforward configuration andmanner of operation.

It is another object of our invention to provide liquid introduction andliquid separation apparatus as above which in each instance, essentiallyrequire no moving parts, and are thus basically maintenance-free.

It is another object of our invention to provide liquid introduction andliquid separation apparatus as above which, in each instance, requirethe use of only readily available, relatively inexpensive materials ofproven effectiveness and dependability to the task at hand, and whichthus may be fabricated at relatively low cost.

It is another object of our invention to provide liquid introduction andliquid separation apparatus and method as above which are not limited byconfiguration or manner of operation to particular locations within aliquid processing system with which the same may be utilized, and whichthus exhibit significant versatility of application.

It is a further object of our invention to provide liquid introductionand liquid separation apparatus and method as above which areparticularly adapted for combined use in a continuous flow, automatedsample liquid analysis system to form an isolation liquidlayer-encapsulated, air-segmented, successive sample liquid stream forthe minimization of sample liquid carryover, and to subsequentlyseparate said sample liquid stream, substantially in whole or in part,from said isolation liquid encapsulating layer to prevent interferencethereof with sample liquid analysis; thereby, in both respectsmaximizing the accuracy of the sample liquid analysis results.

DESCRIPTION OF THE DRAWING

The above and other significant objects and advantages of our inventionare believed made clear by the following detailed description thereoftaken in conjunction with the following drawings wherein:

FIG. 1 is an essentially schematic diagram, with certain parts incross-section, of a sample liquid analysis system including apparatusrepresentatively configured and operable in accordance with theteachings of a first embodiment of our invention for substantiallyseparating at least a portion of one or more liquids out of a flowingstream of immiscible liquid on a continuous flow basis;

FIG. 2 is an enlarged cross-sectional view of the probe means of theapparatus of FIG. 1;

FIG. 3 is an essentially schematic diagram, with certain parts incross-section, of a sample liquid analysis system including apparatusrepresentatively configured and operable in accordance with theteachings of a second embodiment of our invention for substantiallyseparating one or more liquids out of a flowing stream of immiscibleliquids on a continous flow basis;

FIG. 4 is an essentially schematic diagram, of a continuous flow sampleliquid analysis system including the embodiment of the liquid separatingapparatus of our invention of both FIG. 1 and FIG. 3;

FIG. 5 is an essentially schematic diagram, with certain parts incross-section, of a sample liquid analysis system including apparatusrepresentatively configured and operable in accordance with theteachings of a third embodiment of our invention for introducing one ormore liquids into a flowing stream of liquid which is immiscibletherewith to form a flowing stream of said immiscible liquids on acontinuous flow basis and wherein one said liquids substantiallyencapsulates another of said liquids;

FIG. 6 is an essentially schematic diagram, with certain parts incross-section, of a sample liquid analysis system including apparatusrepresentatively configured and operable in accordance with theteachings of a fourth embodiment of our invention for introducing aliquid into a flowing stream of one or more liquids which are immiscibletherewith to form a flowing stream of said immiscible liquids on acontinuous flow basis and wherein one of said liquids substantiallyencapsulates another of said liquids; and

FIG. 7 is an essentially schematic diagram of a continuous flow sampleliquid analysis system including the embodiments of the liquidintroduction apparatus of our invention of FIG. 6, and the liquidseparating apparatus of our invention of FIG. 3.

SUMMARY OF THE INVENTION

As representatively disclosed herein in a first preferred embodiment,the immiscible liquid spearating apparatus and method of our inventioncomprises probe means which are operatively associated with flow conduitmeans to continuously withdraw and thus separate an essentially centralportion of an immiscible liquid stream flowing therethrough. The streamis formed by an air-segmented aqueous sample liquid stream which issubstantially encapsulated in a layer of an immiscible hydrophobicisolation liquid which preferentially "wets" and is disposed at thehydrophobic inner conduit means surface to the substantial exclusion ofthe aqueous sample liquids which are flowing therewithin; thus insuringthat the essentially central portion withdrawn by the probe means issubstantially only aqueous sample liquid. The thusly substantiallyseparated sample liquid stream portion is flowed by pump means from theprobe means to secondary conduit means and therethrough to additionalsample liquid processing and/or analysis means.

As representatively disclosed herein in a second preferred embodiment,the immiscible liquid separating apparatus and method of our inventioncomprise porous flow conduit insert means which are operativelyassociated with flow conduit means to continuously withdraw and thusseparate the outer portion of an immiscible liquid stream flowingtherethrough. This stream is formed by an air-segmented aqueous sampleliquid stream which is substantially encapsulated in a layer of animmiscible hydrophobic isolation liquid which preferentially "wets" andis disposed at the inner conduit means and conduit insert meanshydrophobic surfaces to the substantial exclusion of the aqueous sampleliquids which are flowing therewithin; thus insuring that the outerstream portion withdrawn through the porous conduit insert means issubstantially only the isolation liquid. Sleeve means are provided tosurround the porous conduit insert means and provide a fluid-tight spacetherebetween, and pump means are operatively connected thereto toprovide a differential pressure across the porous conduit insert meansand promote the flow of the isolation liquid therethrough. Theresultant, substantially isolation liquid-free sample liquid stream isflowed through the conduit means downstream of the porous conduit insertmeans to additional sample liquid processing and/or analysis means.

A representative combination in series in accordance with the teachingsof our invention of the first and second preferred embodiments of theimmiscible liquid separation apparatus and method of that invention inthe same sample liquid analysis system for purposes of increase in theoverall liquid separational efficiency is also disclosed.

As representatively disclosed herein in a third preferred embodiment,the liquid introduction apparatus and method of our invention compriseprobe means which are operatively associated with flow conduit means tocontinuously introduce an air-segmented aqueous sample liquid streaminto the essentially central portion of an immiscible hydrophicisolation liquid stream which is flowing in said conduit means, andwhich preferentially "wets" the hydrophobic inner surface of saidconduit means to the substantial exclusion of the aqueous sampleliquids. Introduction of the air-segmented sample liquid stream into theisolation liquid stream through the probe means displaces the latterstream to the inner conduit means surface for retention thereat inaccordance with the preferential "wetting." This effectively forms alayer of the isolation liquid at the inner conduit means surface forencapsulation of the air-segmented, sample liquid stream and continuedflow thereof through the flow conduit means to sample liquid processingand/or analysis means.

As representatively disclosed herein in a fourth preferred embodiment,the liquid introduction apparatus and method of our invention compriseporous flow conduit insert means which are operatively associated withflow conduit means to continuously introduce an immiscible hydrophobicisolation liquid into the outer portion of an air-segmented aqueoussample liquid stream which is flowing in said conduit means. Thisisolation liquid preferentially "wets" the inner hydrophobic surfaces ofsaid flow conduit means and said conduit insert means to the substantialexclusion of the aqueous sample liquids; and thus effectively forms alayer of the isolation liquid at those inner surfaces for encapsulationof the air-segmented, sample liquid stream and continued flow thereofthrough the flow conduit means to sample liquid processing and/oranalysis means.

A representative combination in series in accordance with the teachingsof our invention of the liquid introduction apparatus and method thereofand the liquid spearation apparatus and method thereof to in turn formthe isolation liquid encapsulated air-segmented sample liquid stream andsubsequently substantially separate the sample liquids, or at leastsignificant portions thereof, from the isolation liquid, in the samesample liquid analysis system is also disclosed.

In each instance, it may be readily understood that the formation of theencapsulating isolation liquid layer is to minimize sample liquidcarryover in the sample liquid analysis system, while the substantialseparation of the sample liquids therefrom prior to additional sampleliquid processing and/or analysis is to prevent interference by theisolation liquid with the performance of thse functions. Thus, and inboth events, the accuracy of the sample liquid analysis results ismaximized to significant advantage by the apparatus and method of ourinvention.

DETAILED DESCRIPTION OF THE INVENTION

Referring now to FIG. 1, a first embodiment of apparatusrepresentatively configured and operable in accordance with theteachings of our invention is indicated generally at 10; and comprises aflow conduit 12 of any suitably inert, highly hydrophobic material inthe nature for example of an appropriate fluorinated hydrocarbon.

In addition to being highly hydrophobic, fluorinated hydrocarbonmaterials of the nature utilized in the formation of flow conduit 12 arewell known and understood by those skilled in this art to be selectively"wettable" by a wide range, for example, of fluorinated orperfluorinated hydrocarbon, or silicon liquids of very low surfacetension, which are also hydrophobic, to the substantial exclusion ofaqueous liquids which are immiscible therewith; and this phenomenon ofselective "wettability" of hydrophobic materials by these liquids to thesubstantial exclusion of immiscible aqueous liquids is discussed in somedetail in U.S. Pat. No. 3,479,141 issued Nov. 18, 1969 to William J.Smythe, et al, for "Method and Apparatus For Analysis" and assigned tothe assignee hereof, the disclosure of which is hereby incorporated byreference in this specification.

U.S. Pat. No. 3,479,141 also discloses the use of this concept ofselective "wettability" of hydrophobic materials to minimize aqueoussample liquid carryover, e.g. the contamination of a succeeding aqueoussample liquid by the residue of a preceding aqueous sample liquid,attendant automated, successive aqueous sample liquid analysis. Inaccordance with that patent disclosure, this is accomplished by theencapsulation within an immiscible hydrophobic silicone liquid of thesuccessive aqueous sample liquids of a continuously flowing streamthereof as the same progresses through a fluorinated hydrocarbon flowconduit length; with the silicone liquid functioning as an isolationliquid to isolate the aqueous sample liquids, one from the other, toprevent contact and cross-contamination therebetween, and functioning asan isolation liquid to selectively "wet" the inner flow conduit wall tothe substantial exclusion of the aqueous sample liquids therebyisolating those sample liquids from that inner flow conduit wall andpreventing a preceding aqueous sample liquid from contacting the sameand leaving a residue thereon for pick-up by and contamination of asucceeding aqueous sample liquid

Other United States Patents which relevantly disclose the application ofthis concept of selective "wettability" of hydrophobic conduit materialsby a hydrophobic isolation liquid to the substantial exclusion ofaqueous sample liquids which are immiscible therewith for theminimization of aqueous sample liquid carryover attendant automated,successive aqueous sample liquid analysis are U.S. Pat. No. 4,253,466issued Mar. 3, 1981 to William J. Smythe, et al, for "Method andApparatus For Automated Analysis Of Fluid Samples," and assigned to theassignee hereof, U.S. Pat. Nos. 4,121,466 and 4,357,301 as referred tohereinabove under the "Description of the prior art," and U.S. Pat. No.4,517,302 issued May 14, 1986 to Steven Saros, et al, for "SampleAnalysis System," and assigned to the assignee hereof; and thedisclosures of each of these United States Patents are also incorporatedby reference in this specification.

Sampler and sample processor means are indicated schematically at 14 inFIG. 1: and may, for example, take the general form of those disclosedin U.S. Pat. No. 4,121,466 wherein the same are operable to generate anair-segmented stream 16 of isolation liquid-encapsulated successiveaqueous sample liquid segments in hydrophobic flow conduit 12, therebyminimizing carryover between those aqueous sample liquid segments, andsignificantly increasing the accuracy of the sample liquid analysisresults. In addition, sampler and sample processor means 14 will readilybe understood by those skilled in this art to include any and all sampleliquid processing components as required for the particular sampleliquid analysis of interest; and these may include but not be limited tocomponents for the addition of appropriate reagent(s) to the sampleliquids, and sample liquid dilution, heating, incubation and/or mixingcomponents.

As illustrated in FIG. 1, air-segmented sample liquid stream 16comprises alternating sample liquid segments S1, S2, S3, S4, S5, and S6,as respectively separated in turn by air segments A1, A2, A3, A4, andA5; all as respectively encapsulated as shown by thin layers of theimmiscible isolation liquid as indicated at 20 between the respectivesample liquid and air segments. Although the respective thicknesses ofthe isolation liquid layers 18 and 20 may vary in accordance with theparticular sample liquid analysis application in question, it may beunderstood that, in accordance with the extremely high costs of suitableisolation liquids which may, for example, approach $1,000 per gallon,contemporary versions of the sampler here under discussion have beendeveloped with the capability of generating extremely thin, althoughnonetheless fully effective isolation liquid encapsulating layers 18 and20. In fact, it will be clear to those skilled in this art that naturalspreading and hydrodynamic forces tend to force excess isolation liquidto the inner wall of conduit 12, thus maintaining extremely thininter-segment isolation liquid layers 20, especially at the generallycentral portions thereof, even under relatively "rich" isolation liquidconditions. For example, and with particular regard to the inter-segmentisolation liquid layers 20 of FIG. 1 which are not in contact with thehighly hydrophobic inner wall of flow conduit 12, generation of suchlayers of essentially monolayer or single molecule thickness, at leastat the generally central portions thereof, is now believed to bepossible.

Liquid separator means are indicated generally at 22 in FIG. 1 andcomprise a generally L-shaped tubular probe 23 including a probe length24 which extends as shown through the wall of flow conduit 12, and aconnected probe length 26 which extends therefrom within flow conduit 12concentrically thereof in the upstream direction with regard toair-segmented sample liquid stream 16. Sealing means are indicated at 28and are disposed at the juncture of probe length 24 and flow conduit 12to prevent leakage from the flow conduit at that juncture.

As best seen in FIG. 2, probe length 26 includes a sharp-edged inlet end30 communicating the internal probe flow passage 32 with the airsegmented sample liquid stream 16 in flow conduit 12.

Probe 23 is fabricated from any appropriately inert, hydrophilicmaterial, for example stainless steel as shown, or glass; and, as bestseen in FIG. 2, the exterior of probe 23 which is exposed to the airsegmented sample liquid stream 16 within flow conduit 12 is surfacecoated with an appropriately inert, highly hydrophobic material in thenature, for example of a fluorinated hydrocarbon, as indicated at 34.Under these circumstances, it will be clear to those skilled in this artthat the isolation liquid will be preferentially attracted to the highlyhydrophobic surface coating 34 at the exterior of probe 23 to "wet" thesame to the substantial exclusion of the aqueous sample liquids, whilethe aqueous sample liquids will in turn be preferentially attracted tothe hydrophilic wall of the internal probe flow passage 32 to "wet" thesame; and the resultant essentially natural capability of the probe 23to effectively discriminate between the aqueous sample liquids and theimmiscible isolation liquid, especially at the sharp-edged probe inlet30, will be seen to be of particular significant advantage with regardto the effective separation of those liquids as described in detailhereinbelow.

Although dimensions may of course vary in accordance with the particularapplication(s) to which the apparatus of our invention is put, it may beunderstood that with a flow conduit 12 of nominally 1.50 mm innerdiamenter, a representative outer diameter for probe lengths 24 and 26would be nominally 0.75 mm, with a nominal diameter for internal probepassage 32 of 0.25 mm, to thus provide a probe length wall thickness ofnominally 0.25 mm. Under these conditions, a representative nominallength for probe length 26 would be 1.50 cm.

For reasons made clearer hereinbelow, it is of particular importancethat the probe length 26 be centered as precisely as possible vis-a-visflow conduit 12 to insure that the probe intercepts the most central,and thus lowest isolation liquid volume-bearing, portion of sampleliquid stream 16; and that the probe length 26 be of the smallestpractical wall thickness to insure minimal adverse effect thereof on thehydrodynamics of the sample liquid stream 16 at the juncture thereof.

Additional sample liquid processing and/or sample liquid analysis meansare indicated schematically at 36 in FIG. 1 and may take any appropriateform(s) compatible with automated successive sample liquid processingand/or analysis. Most relevantly with regard to the apparatus of ourinvention, and for applications thereof to the automated successiveprocessing and quantitative analysis in turn of a series of bloodsamples, the additional sample liquid processing means may, for example,comprise a dialysis device; while the sample liquid analysis means may,for example, comprise colorimetric, spectroscopic, ion selectiveelectrode, cell counting, enzyme coil and/or dry chemistry slideanalytical devices, wherein the continued presence of the isolationliquid with the sample liquids can interfere with the requisite highlyprecise operation of those analysis devices with resultant, andcontemporarily unacceptable, degradation in the accuracy of the sampleanalysis results. For application of the apparatus of our inventionwherein no additional processing of the aqueous sample liquids from flowconduit 12 is required prior to the automated successive analysisthereof, means 36 would of course be limited to sample analysisdevice(s).

A secondary flow conduit is shown at 38 in FIG. 1, and is connected asindicated through pump 39 to the additional sample liquid processingand/or sample liquid analysis means 36 to supply sample liquids theretoin turn at a flow rate Q2 in accordance with the output of pump 39.Although representatively depicted as of essentially the same dimensionas flow conduit 12, it will be clear to those skilled in this art thatsecondary flow conduit 38 may be of any dimension and compositioncompatible with additional sample liquid processing and/or analysis, forexample standard laboratory tubing. With secondary flow conduit 38 ofessentially the same dimension as flow conduit 12, and thus ofsignificantly greater outer diameter than probe length 24, anessentially L-shaped connecting piece 40, including an enlarged endportion 42, of essentially the same size as secondary flow conduit 38,is provided to connect the outlet end of probe length 24 to the inletend of secondary flow conduit 38 as shown, and of course in fluid-tightmanner. Alternatively, connecting piece 40 may, for example, be formedintegrally with probe 23.

Isolation liquid recovery means are indicated schematically at 44 inFIG. 1 and may take any form compatible with the effective recovery ofthe isolation liquid, which may be quite expensive as set forthhereinabove, from the sample liquids, for re-use of the isolationliquid. Since, in the vast majority of instances, the isolation liquidwill be of different specific gravity than the sample liquids, therecovery of the isolation liquid may be effectively accomplished inunhurried manner by simply allowing the respective sample and isolationliquids to settle out at different levels under the force of gravity inthe same container, and subsequently removing the isolation liquidtherefrom for re-use.

Flow conduit 12 is connected as indicated in FIG. 1 to isolation liquidrecovery means 44 to supply the isolation liquid-rich sample liquidstream 16 thereto.

A system controller is indicated schematically at 48 in FIG. 1, and mayfor example take the form of an appropriately programmablemicroprocessor device. Controller 48 is electrically connected asindicated by lines 50, 52 and 54 to sampler and sample processing means14, pump 39, and additional sample processing and/or sample analysismeans 36 to control and synchronize the respective operations thereof.

In operation of the apparatus 10 for the removal of substantiallyisolation liquid free sample liquid and air segment portions fromflowing air segmented sample liquid stream 16 in flow conduit 12, andthe supply of the same in turn in the form of a substantially isolationliquid free, air segmented sample liquid stream as indicated at 58 inFIG. 1 in secondary flow conduit 38 to additional sample liquidprocessing and/or analysis means 36 for successive analysis in turn ofeach of those sample liquid segment portions, it may be understood thatpump 39 is preferably operated by controller 48 at a flow rate Q2 whichis slightly less than the flow rate Q1 at which the air segmented sampleliquid stream 16 is flowed in flow conduit 12 immediately upstream ofthe inlet edge 30 of the probe means 23. A representative example of theratio Q2/Q1 is 0.90, whereby will be clear that nearly all of the sampleliquid stream 16 will be withdrawn from flow conduit 12 by probe means23 for flow as sample liquid stream 58 in secondary flow conduit 38.Under these conditions, the flow rate of the remainder of sample liquidstream 16 in flow conduit 12 downstream of probe means 23 to theisolation liquid recovery means 44 will as indicated be equal to Q1-Q2.Of course, it will be immediately clear to those skilled in this artthat pumping arrangements other than that shown, for example,differential pumping or pressure pumping, may be utilized to achieve thedesired ratio between Q2 and Q1.

FIG. 2 makes clear that as each inter-segment isolation liquid layer 20in air segmented sample liquid stream 16 is flowed into contact with thesharp inlet edge 30 of probe means 23, that isolation liquid layer willbe cleanly cut by the same, with only the particularly low isolationliquid volume of the most central portion of that isolation liquid layeras indicated at 20C in FIG. 2 being withdrawn along with the sampleliquid from the sample liquid segment of interest as shown into theinternal probe means flow passage 32 for flow as described into andthrough secondary flow conduit 38.

Of particular significance in accordance with the teachings of ourinvention are the facts that the remainder of the isolation liquid layer20 of interest, e.g. that portion thereof disposed radially outward ofprobe means inlet edge 30 and which will most probably re-form in eachinstance as shown in FIG. 1 into a very thin isolation liquid layer 20D,and all of the vastly greater isolation liquid volume bearing layer 18which resides primarily at the inner wall of flow conduit 12, and whichwill thus be greatly "enriched" vis-a-vis the reduced volume sampleliquid segment of interest in each instance as shown at 18D in FIG. 1,will respectively remain in sample liquid stream 16 downstream of theprobe means inlet edge 30 for flow as described therewith to theisolation liquid recovery means 44; and it will be immediately clear tothose skilled in this art that the highly hydrophobic surface coating 34on the external surface of probe means 23, and the highly hydrophobicinner wall surface of flow conduit 12--both of which operate asdescribed to strongly attract the isolation liquid to the substantialexclusion of the aqueous sample liquids--will also operate toparticularly significant advantage in this regard. Of course, thehydrophilic surface of the internal probe means flow passage 32 whichselectively attracts the aqueous sample liquids is also of significantadvantage in this regard.

In accordance with the above, the apparatus 10 of our invention will beunderstood to operate to provide a substantially isolation liquid freeair-segmented sample liquid stream 58 in secondary flow conduit 38 forflow as described to additional sample liquid processing and/or analysismeans 36 for highly accurate, automated successive sample liquidanalysis; it having been determined by actual tests that the extremelysmall amounts of isolation liquid 20C which do remain in sample liquidstream 58, and which will most probably simply disperse throughout therespective sample liquid segments thereof as shown in FIG. 1, will, inthe vast majority of instances, be of no clinical significance withregard to adverse effect upon the overall efficiency and/or accuracy ofthe operation of the additional sample liquid processing and/or sampleliquid analysis means 36.

A second embodiment of apparatus representatively configured andoperable in accordance with the teachings of our invention is indicatedgenerally at 60 in FIG. 3; and comprises a flow conduit 62, again of anysuitably inert, highly hydrophobic material in the nature of anappropriate fluorinated hydrocarbon.

Liquid separator means are indicated generally at 63 in FIG. 3 andcomprise a conduit insert 64 which is preferably of the same innerdiameter as flow conduit 62, and which is joined therein and attachedthereto as shown in fluid-tight manner by, for example, a suitableadhesive as indicated at 66 in FIG. 3. Insert 64 is fabricated from anysuitably porous filter material which is also highly hydrophobic, againfor example an appropriate fluorinated hydrocarbon.

A generally coaxial sleeve is indicated at 68 and is secured as shown influid-tight manner to the adjacent external walls of flow conduit 62,again for example by a suitable adhesive as indicated at 70, to surroundconduit insert 64 and provide a generally torous-like enclosed space 72therebetween. Sleeve 68 is made of any suitable, fluid-tight material,for example a suitable plastic. Annular seals 74, 76, 78 and 80 arerespectively provided as shown at the junctures of the insert 64 and thesleeve 68 with the flow conduit 62 to prevent fluid leakage therefrom.Alternatively, these junctures may be formed by fluid-tight press fits.

A secondary flow conduit is indicated at 82 in FIG. 3 and extends asshown through porous insert 64 at the lower portion thereof intocommunication with enclosed space 72. A fluid-tight annular seal 83, ora like press fit, is utilized to prevent fluid leakage at this juncture.Secondary flow conduit 82 is connected as indicated through pump 84 tosupply isolation liquid from space 72 to isolation liquid recovery meansas again indicated schematically at 44, and which may take the same formas described hereinabove with regard to the embodiment of FIG. 1.

Sampler and sample processor means, additional sample processing, and/orsample analysis means, and system controller are respectively againindicated schematically at 14, 36 and 48; and, in each instance, maytake the same form as described hereinabove with regard to theembodiment of FIG. 1.

In the operation of apparatus 60 of FIG. 3, sampler and sample processormeans 14 are again operable as described to generate an air-segmentedstream as there indicated at 86 of isolation liquid encapsulatedsuccessive aqueous sample liquid segments in flow conduit 62; and thisstream 86 will be seen in FIG. 3 to comprise alternating aqueous sampleliquid segments S1, S2, S3, and S4, as respectively separated in turn byair segments A1, A2, and A3; all as respectively encapsulated as shownby thin layers of the isolation liquid as again indicated at 18 betweenthe respective sample liquid and air segments and the inner wall of flowconduit 62, and as again indicated at 20 between the respective sampleliquid and air segments.

With air-segmented sample liquid stream 86 flowing as described in flowconduit 63 at a pressure P1, and with pump 84 operated by controller 48to create a lower pressure P2 in sleeve space 72 through secondary flowconduit 82, a differential pressure PD is created across the poroushydrophobic conduit insert 64. As a result, as each of the respectiveaqueous sample liquid segments S and air segments A flows into insert64, this differential pressure PD taken in conjunction with theinterfacial tension driving forces arising from the selective attractionof the isolation liquid to the highly hydrophobic material of the insert64, will operate to draw the isolation liquid layers 18 at the insertwall essentially in their respective entireties and, to a somewhatlesser extent, the inter-segment isolation liquid layers 20, through theporous hydrophobic insert 64 into the sleeve space 72 as shown in FIG. 3thereby separating the same from the segmented sample liquid stream 86.With more specific regard to the inter-segment isolation liquid layers20, it will be clear that the same will, in effect, be naturally drawnto the inner wall of porous insert 64 upon the removal of the isolationliquid layer 18 for removal from the sample liquid stream 86therethrough as a result of spreading and hydrodynamic forces; with theremaining very small portions of those inter-segment layers 20 which arenot so drawn most probably simply dispersing throughout the sampleliquid segment as indicated at 20C in FIG. 3 in and downstream of theinsert 64. The high interfacial tension between the aqueous sampleliquid segments and the porous hydrophobic insert 64 will operate tosubstantially inhibit aqueous sample liquid passage through insert 64.

Under the above circumstances, it will be clear to those skilled in thisart that proper selection of the differential pressure PD and the poresize of the porous insert 64, it becomes possible to withdrawsubstantially only isolation liquid, plus some measure of air from therespective air segments A which is of no clinical significance, from theair-segmented sample liquid stream 86 into sleeve space 72 as described;while appropriate selection of the length of the insert 64 in accordancewith the above can be utilized to insure that the maximum quantity ofthe isolation liquid which can, as a practical matter, be withdrawn fromthe stream 86 will, in fact, be withdrawn therefrom attendant thetransit by the stream of the insert 64. Although these controllingparameters may, of course, vary in accordance with the particularapplication(s) to which the apparatus of our invention is put, valuesthereof for a representative application of that apparatus with aninsert of nominally 1.50 mm inner diameter and a nominal length of 3.0cm, would be a nominal pore size of 2.0 microns for the porous insert64, and a nominal pressure differential PD equal to P1-P2 across theinsert 64 of up to 10 psi to provide for the withdrawal of substantiallyonly the isolation liquid and some measure of air as described throughthe porous insert 64 into sleeve space 72.

For application of the apparatus 60 wherein the maximum achievableremoval of the isolation liquid from the air-segmented sample liquidstream 86 is required, and wherein some relatively small loss of theaqueous sample liquids through porous insert 64, for example 5.0% byvolume, can be accommodated without adverse effect upon the accuracy ofthe sample liquid analysis results, it will be clear that the pore sizeof the porous insert 64, the differential pressure PD, and/or the lengthof porous insert 64 can be increased to that effect.

FIG. 3 makes clear that the isolation liquid which is withdrawn asdescribed through porous insert 64 into space 72, plus any air and/oraqueous sample liquids as may be included therewith, will be pumped bypump 84 to isolation liquid recovery means 44 for isolation liquidrecovery and re-use, if desired, as described herein above with regardto the embodiment of FIG. 1. Concomitantly, the now substantiallyisolation liquid-free air-segmented sample liquid stream 86, includingsubstantially only the remaining very low isolation liquidvolume-bearing isolation liquid layer portions 20C, will exit sleeve 64for flow as indicated through conduit to additional sample processingand/or sample analysis means 36 for highly accurate, automatedsuccessive aqueous sample liquid processing and/or analysis; it havingalso been determined by the actual test of the apparatus 60 of ourinvention that the extremely small amounts of isolation liquid 20C whichdo remain as described in the sample liquid stream 86 will, in the vastmajority of instances, be of no clinical significance with regard toadverse effect upon the operation of the additional sample liquidprocessing and/or analysis means 36.

For certain applications of the apparatus and method of our inventionwherein an absolute maximum of achievable isolation liquid stream isrequired prior to additional sample liquid processing and/or analysis,the liquid separator means of our invention may be serially connected orcascaded. FIG. 4 schematically depicts a representative apparatusarrangement to that effect wherein the liquid separator means 22 of FIG.1 and the liquid separator means 63 are serially connected in thatorder. In operation, it will be clear that the substantially isolationliquid-free air-segmented sample liquid stream 58 from secondary flowconduit 38 of FIG. 1 would be flowed in turn as air-segmented sampleliquid stream 86 through flow conduit 62 of FIG. 3 by pump 39 foradditional isolation liquid removal therefrom as described within sleeve68 of FIG. 3;. it being noted in this regard that the very smallisolation liquid quantities initially remaining in stream 58 as formedby the central portions 20C of isolation liquid layers 20 will mostprobably soon migrate within secondary flow conduit 38 to the inner wallthereof--which, in this instance, would of course be made from a highlyhydrophobic material--to again provide for significant inhibition ofsample liquid carryover therein, and to enable the subsequent, highlyeffective removal thereof from the flowing sample liquid stream 86 inflow conduit 62 by the withdrawal thereof as described through porousconduit insert 64 within sleeve 68 as described. In this instance, flowconduit 12 and secondary flow conduit 82 would be connected as indicatedto the same isolation liquid recovery means 44.

As alternatives to the serial liquid spearator means arrangement of FIG.4, it will be clear to those skilled in this art that the same could beserially connected with the liquid separator means 63 of FIG. 3 ahead ofthe liquid separator means 22 of FIG. 1, and that two or more of thesame liquid separator means 22 or 63 could be serially connected assuch; again in any event to improve the overall efficiency of theisolation liquid removal process.

Although disclosed in detail hereinabove as applied to the particularlyeffective substantial separation of the isolation liquid from anisolation liquid encapsulated, segmented sample liquid stream prior tosample liquid analysis to improve the accuracy thereof, it will be clearthat the embodiments of the liquid separating apparatus 10 and 60 of ourinvention can alternatively be applied to that purpose solely forisolation liquid reclamation following sample analysis in thoseinstances wherein the sample liquid analysis can be satisfactorilyperformed with the isolation liquid layers essentially in place asdescribed. In such applications, the substantially isolation liquid-freesample liquid streams downstream of the separating apparatus would mostprobably simply be flowed to waste.

A third embodiment of apparatus representatively configured and operablein accordance with the teachings of our invention is indicated generallyat 90 in FIG. 5; and comprises a flow conduit 92, again of any suitablyinert, highly hydrophobic material in the nature of an appropriatefluorinated hydrocarbon.

An isolation liquid source is indicated schematically at 94, and isconnected as shown through pump 96 to supply a stream 98 of theisolation liquid for flow through the conduit 92 in the directionindicated.

Liquid introduction means are indicated generally at 100 in FIG. 5, andcomprise a generally L-shaped tubular probe 102 including a probe length104 which extends as shown through the wall of flow conduit 92, and aconnected probe length 106 which extends therefrom within flow conduit92 concentrically thereof in the upstream direction with regard toisolation liquid stream 98. Sealing means are indicated at 108 and aredisposed at the juncture of probe length 104 and conduit 92 to preventfluid leakage at that juncture. Probe 102 includes an internal flowpassage 110 extending therethrough as shown; and is fabricated from anyappropriately inert material, for example stainless steel as shown, orglass.

Although dimensions may, of course, vary in accordance with theparticular application(s) to which the apparatus of our invention isput, it may be understood that with a flow conduit 92 of nominally 1.50mm inner diameter, a representative outer diameter for probe lengths 104and 106 would be nominally 1.3 mm, with a nominal diameter for internalprobe flow passage 110 of 1.0 mm, to thus provide probe wall thicknessesof nominally 0.15 mm. Under these conditions, a representative nominallength for probe length 106 would be 1.50 cm. For reasons made clearerhereinbelow, it is again of particular importance that probe length 106be centered as precisely as possible vis-a-vis flow conduit 92 to insurethat this probe length lies in the most central portion of the isolationliquid stream 98 flowing therearound. Minimal probe wall thickness isalso again of importance.

Sampler means are indicated schematically at 112 in FIG. 5 and areoperable in conventional manner to supply an air-segmented stream 114 ofsample liquid segments as shown to probe 102 for flow therethroughthrough internal probe flow passage 110 to merge with the isolationliquid stream 98 as the stream 114 exits the outlet edge 116 of theprobe length 106. As illustrated in FIG. 5, stream 114 comprises sampleliquid segments S1 and S2 and air segments A1 and A2.

In operation, FIG. 5 makes clear that as each air segment, for exampleair segment A1 as shown, exits the internal probe flow passage 110through outlet edge 116 of the probe length 106, that air segment willnaturally expand in diameter to essentially fill the larger diameterinternal flow passage 118 of the conduit 92 and, in so doing, thesegment or bubble meniscus 120 will pick up the flowing isolation liquidfrom isolation liquid stream 98 and displace and distribute the same inthe form of the isolation liquid layer 18 at the wall 118 that internalconduit flow passage to coaxially surround the air segment of interest.

The immediately following flow of the succeeding sample liquid segmentS1 from the internal probe flow passage 110 into the isolation liquidstream 98 in internal flow conduit passage will, as that sample liquidsegment naturally expands in diameter in like manner to essentially fillthe larger diameter flow passage 118 of the conduit 92, continue thisdisplacement and distribution of the isolation liquid, therebycontinuing the formation of the isolation liquid layer 18. Of course,the preferential "wettability" of the highly hydrophobic wall of theinternal conduit flow passage 118 by the isolation liquid to thesubstantial exclusion of the aqueous sample liquids virtually insuresagainst any contact by the aqueous sample liquids with that internalconduit flow passage wall attendant the flow of the sample liquid stream114 into conduit 92 as described. This, in turn, virtually insures theformation and retention in conduit 92 downstream of probe 102 of acontinuous, air and sample liquid stream encapsulating isolation liquidlayer 18. In addition, and as each air segment, for example A1, exitsthe probe 102 for expansion as shown in FIG. 5 within the isolationliquid stream 98 in conduit 92, the same will also operate to displacethe isolation liquid from that stream to continue the formation of theisolation liquid layer 18, and to form the inter-sample liquid segmentisoaltion liquid layers 20 as shown in FIG. 5 at the respectiveair-sample liquid segment interfaces.

In accordance with the above, the resultant isolationliquid-encapsulated, air-segmented sample liquid stream 120 in conduit92 in FIG. 5 downstream of probe length 106 and, which as illustratedcomprises sample liquid segments S3 and S4, and air segment A3, will befully protected against sample liquid carryover by the isolation liquidlayer.

Sample liquid processing and/or analysis means are indicatedschematically at 122 in FIG. 5, and are operable as indicated to receivethe isolation liquid encapsulated, air-segmented sample liquid stream120 from conduit 92 to those purposes. In this instance, sample liquidprocessing and/or analysis means 122 would take the form of those whichare satisfactorily operable to those purposes without degradation ofclinical significance in sample liquid processing and/or analysis withthe isolation liquid layer 18 in place.

A system controller is again indicated schematically at 48 in FIG. 5,and is operable as indicated through lines 124, 126 and 125 tosynchronize and control the operation of pump 96, sampler 112, andsample processing and/or analysis means 122.

With pump 96 supplying the isolation liquid stream 198 to flow conduit92 at a flow rate Q1, and sampler 112 supplying the air-segmented sampleliquid stream 114 to flow conduit 92 at a flow rate Q2, it will be clearthat the flow rate Q3 of stream 120 in flow conduit 92 will be equal toQ1 and Q2. Although these flow rates, and the ratio between flow rate Q2and Q1 may, of course, vary throughout wide ranges in accordance withthe application(s) to which the apparatus of 90 of FIG. 5 are put, itmay be understood that a representative range for the ratio Q2/Q1 isbetween 10 to 1 and 100 to 1 in accordance with the required "richness"of the isolation liquid layer 18 relative to stream 120.

Of particular advantage with regard to the apparatus 90 of FIG. 5 arethe facts that the same operates to precisely form the isolation liquidlayer 18 where it is most needed and most effective for purposes ofminimization of sample liquid carryover, e.g. at the inner wall 118 offlow conduit 92, and that precise and readily accomplishable control ineach instance of the flow rate ratio Q2/Q1 enables the precise formationof the isolation liquid layer of the optimal thickness in accordancewith the particular application to which the apparatus 90 are to be putwith regard to both sample liquid carryover minimization and economy ofisolation liquid utilization.

A fourth embodiment of apparatus representatively configured andoperable in accordance with the teachings of our invention is indicatedgenerally at 130 in FIG. 6; and comprises a flow conduit 132, again ofany suitably inert, highly hydrophobic material in the nature of anappropriate fluorinated hydrocarbon.

A sampler is indicated schematically at 134 and is operable as indicatedin conventional manner to supply an air-segmented sample liquid stream136 for flow through the internal flow passage 138 of conduit 132. Asillustrated in FIG. 6, stream 136 comprises sample liquid segments S1,S2, S3 and S4 as separated by air segments A1, A2 and A3.

Liquid introduction means are indicated generally at 138 in FIG. 6, andtake much the same form structurally as the liquid separator means 63 ofFIG. 3. To that effect, liquid introduction means 138 comprise a conduitinsert 140 which is preferably of the same inner diameter as flowconduit 132, and which is joined therein and attached thereto as shownin fluid-tight manner by, for example, a suitable adhesive as indicatedat 142 in FIG. 6. Insert 140 is fabricated from any suitably porousfilter material which is also highly hydrophobic, again for example anappropriate fluorinated hydrocarbon.

A generally coaxial sleeve is indicated at 144 and is secured as shownin fluid-tight manner to the adjacent external walls of conduit 132,again for example by a suitable adhesive as indicated at 146, tosurround the conduit insert 144 and provide a generally torous-likeenclosed space 148 therebetween. Sleeve 144 is made of any suitable,fluid-tight material, for example a suitable plastic. Annular seals 150,152, 154 and 156 are provided as shown at the respective junctures ofthe insert 140 and sleeve 144 with the flow conduit 132 to prevent fluidleakage therefrom. Again, fluid-tight press fits may alternatively beutilized to form these junctures.

An isolation liquid supply conduit is indicated at 160 in FIG. 6 andextends as shown through sleeve 144 into communication with torous-likespace 148. An annular seal 162 is disposed around the juncture ofconduit 160 and sleeve 144 to prevent fluid leakage therefrom.

An isolation liquid source is indicated schematically at 164 in FIG. 6and is connected as shown through a pump as schematically indicated at166 for the supply of isolation liquid through supply conduit 160 to thespace 148. Pump 166 is preferably of the positive displacement typetaking, for example, the form of a standard laboratory syringe pumpwhich is operable to positively pump liquids at precisely determinableflow rates.

In operation of the apparatus 138 of FIG. 6 to introduce an isolationliquid from source 164 to air-segmented sample liquid stream 136 withininsert 140 to encapsulate the latter therewithin, it will be clear that,with torous-like space 148 filled with the isolation liquid, thecontinued operation of positive displacement pump 166 will function toflow the isolation liquid from space 148 through the porous insert 140to emerge at the inner wall 168 of the same at the outer boundary of theflowing air segmented sample liquid stream 136 therein. Of course, asthe positively pumped isolation liquid emerges from the inner wall 168of the insert 140, it will displace the air-segmented sample liquidstream 136 therefrom; with the preferential attraction of the isolationliquid to that highly hydrophobic inner wall to the substantialexclusion of the aqueous sample liquids, functioning to retain theisolation liquid in contact therewith to form the isolation liquid layer18 within insert 140 for flow with the stream 136 through flow conduit132 in the direction indicated in FIG. 6. In addition, and with asufficiently "high" ratio of isolation liquid flow Q1 through porousconduit insert 140 to air-segmented sample liquid stream flow Q2 throughconduit 132, formation of the respective isolation liquid layers 20 atthe respective air and sample liquid segment interfaces as shown in FIG.6 will also again be accomplished. As a result, it will be clear thatsample liquid carryover is again effectively minimized with regard tothe isolation liquid-encapsulated, air-segmented sample liquid stream174 attendant the continued flow thereof in flow conduit 132 downstreamof the porous conduit insert 140.

Sample processing and/or analysis means are schematically indicated at176 in FIG. 6, and operate as indicated to receive the isolation liquidencapsulated, air-segmented sample liquid stream 172 from flow conduit132 to those purposes; it being clear that, in this instance, sampleliquid processing and/or analysis means 176 would take the form of thosewhich are satisfactorily operable without degradation of clinicalsignificance in sample liquid processing and/or analysis with theisolation liquid layers 18 and 20 in place.

A system controller is indicated schematically at 178 in FIG. 6 and isoperatively connected as indicated to sampler 134, pump 166, and sampleprocessing and/or analysis means 176 to control and synchronize therespective operations thereof.

With pump 166 supplying the isolation liquid stream through porousinsert 140 to flow conduit 132 at a flow rate Q1, and sampler 134supplying the air-segmented sample liquid stream 136 to that flowconduit at a flow rate Q2, it will be clear that the flow rate Q3 ofstream 172 will be equal to Q1 and Q2. Again, these flow rates, and theratio between Q1 and Q2 may, of course, vary widely .as set forthhereinabove with regard to the embodiment of FIG. 5.

Again of particular advantage with regard to the apparatus 130 of FIG. 6are the facts that the same operates to precisely form the isolationliquid layer 18 where it is most needed and most effective for purposesof minimization of sample liquid carryover, e.g. at the inner wall 168of the insert 140 and thus at the inner wall 174 of the flow conduit132, and that precise and readily accomplishable control in eachinstance of the flow rate ratio Q2/Q1 enables the precise formation ofthe isolation liquid layer 18 of the optimal thickness in accordancewith the particular application to which the apparatus 130 are to be putwith regard to both sample liquid carryover minimization andminimization of isolation liquid consumption.

FIG. 7 illustrates a sample analysis system as generally indicated at190 and which includes both the liquid introduction apparatus 130 ofFIG. 6, and the liquid separation apparatus 60 of FIG. 3. Sampler,isolation liquid source, and additional sample processing and/or sampleanalysis means, are respectively schematically indicated at 134, 164 and36; while a system controller is schematically indicated at 192, and isconnected as shown by lines 180, 182, 52 and 54 to sampler 134, pump166, pump 84 and additional sample processing and/or sample analysismeans 36 to control and synchronize the respective operations thereof.Sample processing means are indicated schematically at 194, and areoperatively disposed as shown between liquid introduction apparatus 130and liquid separation apparatus 60. A flow conduit 196 is provided asshown in the system of FIG. 7 to connect the outlet of pump 84 to theisolation liquid source 164 to return the separated isolation liquid tothe latter.

In the operation of the sample analysis system 190 of FIG. 7, anair-segmented sample liquid stream is supplied from sampler 134 throughflow conduit 132 to liquid introduction apparatus 130 for encapsulationof that stream in an isolation liquid layer 18 as described in detailhereinabove with regard to FIG. 6. This stream then flows as indicatedto and through sample processing means 194 for processing as required,and therefrom as indicated to and through liquid separation apparatus 60for removal of the isolation liquid layer therefrom as described indetail hereinabove with regard to FIG. 3, with the thusly removedisolation liquid being returned as indicated from pump 84 throughconduit 196 to isolation liquid source 164 for continuous re-use in thesystem 190. The resultant, substantially isolation liquid-free,air-segmented sample liquid stream is then flowed as indicated throughconduit 62 to additional sample processing and/or sample analysis means36 to those purposes.

Under the above circumstances, it will be clear that the sample analysissystem 190 of FIG. 7 provides the particularly significant combinedadvantages of precise isolation liquid layer formation for maximalsample liquid carryover prevention, and highly effective removal of thatisolation liquid layer prior to additional sample liquid processingand/or analysis to maximize the effectiveness and accuracy of the same.In fact, since encapsulation of the air-segmented sample liquid streamby the apparatus 130 is essentially limited to the formation of theisolation liquid layer 18 at the inner wall of flow conduit 132, andsince apparatus 60 are effective to remove substantially all of thatisolation liquid layer, it will be clear that the air-segmented sampleliquid stream which is flowed as described from apparatus 60 toadditional sample processing and/or sample analysis means 36 throughflow conduit 62 in FIG. 6 will be virtually isolation liquid-free. Too,since the system 190 functions to constantly recirculate and re-useessentially the same isolation liquid, it will be clear that maximumeconomy of isolation liquid utilization is, of course, achieved.

Although representatively depicted and described as utilizing theapparatus 130 of FIG. 6 for formation of the isolation liquidencapsulated, air-segmented sample liquid stream, and the apparatus 60of FIG. 3. for the separation of the isolation liquid from that sampleliquid stream, it will be immediately clear to those skilled in this artthat the sample analysis system 190 of FIG. 7 could alternativelyutilize the apparatus 90 of FIG. 5 for isolation liquid encapsulatedsample liquid stream formation, and/or the apparatus 10 of FIG. 1 forseparation of the isolation liquid from that stream.

Although disclosed hereinabove by way of representative examples asoperable in conjunction with air-segmented sample liquid streams, itwill be clear that the apparatus and method of our invention are by nomeans limited to use with sample liquids; and that, with the exceptionof the apparatus 100 of FIG. 5 wherein the expanding meniscuses of theair segments operate as described to displace the isolation liquid tothe inner wall 118 of flow conduit 92, the apparatus and method would beoperable to equally advantageous effect with non-air segmentedcontinuous liquid streams.

Various changes may of course be made in the herein disclosed preferredembodiments of the apparatus and method of our invention withoutdeparting from the spirit and scope thereof as defined by the appendedclaims.

What is claimed is:
 1. In apparatus for the substantial separation of asecond liquid from a common stream of said second liquid and a firstliquid which is immiscible therewith flowing in conduit means, saidconduit means including an inner surface which is selectively wettableby said second liquid to the substantial exclusion of said first liquid,and wherein said second liquid is flowing primarily at said innerconduit means surface in substantial contact therewith, and said firstliquid is flowing primarily within said second liquid substantially outof contact with said conduit means inner surface, the improvementscomprising, liquid separating means in said conduit means forsubstantially separating said second liquid from said common stream ofsaid first and second immiscible liquids attendant the flow thereof pastsaid liquid separating means in said conduit means, and meansoperatively connected to said liquid separating means for flowing thethusly separated second liquid out of said conduit means while enablingthe continued flow of the remainder of said common liquids streamthrough said conduit means, said liquid separating means comprising aporous conduit means portion of a material which is selectively wettableby said second liquid to the substantial exclusion of said first liquid,said porous conduit means portion being disposed in said conduit meansso as to be initially contacted substantially only by said second liquidattendant the flow of said first and second immiscible liquids throughsaid conduit means, and means operatively associated with said porousconduit means portion for flowing said second liquid therethrough out ofsaid conduit means.
 2. In apparatus as in claim 1, the improvementsfurther comprising, said porous conduit means portion comprising aconduit means insert which is of essentially the same inner diameter assaid conduit means.
 3. In apparatus as in claim 2 wherein said innerconduit means surface and said porous conduit means insert arehydrophobic, said second liquid is an isolation liquid which ispreferentially attracted thereto to the substantial exclusion of aqueousliquids, and said first liquid is an aqueous sample liquid which isimmiscible with said isolation liquid, the improvements furthercomprising, sample liquid analysis means operatively connected to saidconduit means downstream therein of said porous conduit means insert andoperable to analyze said first liquid.
 4. In apparatus as in claim 1,the improvements further comprising, said means for flowing said secondliquid through said porous conduit means portion out of said conduitmeans comprise, means to create a differential pressure across saidporous conduit means portion.
 5. In apparatus as in claim 1, theimprovements further comprising, sleeve means surrounding said porousconduit means portion externally of the same to create a liquid-tightspace therebetween for the collection of said second liquid upon theflow thereof through said porous conduit means portion.
 6. In apparatusas in claim 5, the improvements further comprising, said means to flowsaid second liquid through said porous conduit means portion comprising,pump means operatively connected to said liquid-tight space and operableto create a pressure therein which is lower than the pressure in saidconduit means.
 7. In liquid separating apparatus for substantiallyseparating at least a portion of a first liquid from a common liquidsstream of first and second immiscible liquids flowing in conduit meansincluding an inner conduit means surface which is selectively wettableby said second liquid to the substantial exclusion of said first liquid,and wherein said first liquid is flowing primarily within said secondliquid substantially out of contact with said conduit means innersurface, the improvements comprising, liquid separating means in saidconduit means for substantially separating at least a portion of saidfirst liquid from said common liquids stream attendant the flow thereofpast said liquid separating means in said conduit means, and meansoperatively connected thereto for flowing the thusly substantiallyseparated first liquid portion out of said conduit means while enablingthe continued flow of the remainder of said common liquids streamthrough said conduit means, said liquid separating means comprisingprobe means including a probe means portion which extends within saidconduit means out of contact with said inner conduit means surface tointercept substantially only the flow of said first liquid in saidconduit means, said probe means portion comprising inlet means forliquid flow thereinto, said means for flowing the thusly substantiallyseparated first liquid portion out of said conduit means comprisinganother portion of said probe means in communication with saidfirst-mentioned probe means portion and extending through the wall ofsaid conduit means for the flow of the thusly substantially separatedfirst liquid portion therethrough out of said conduit means, andsecondary conduit means operatively connected to said another probemeans portion for the flow of the thusly substantially separated firstliquid portion therethrough.
 8. In apparatus as in claim 7 wherein, saidinner conduit means surface is hydrophobic, said second liquid is anisolation liquid which is preferentially attached thereto to thesubstantial exclusion of aqueous liquids and said first liquid is anaqueous sample liquid which is immiscible with said isolation liquid,the improvements further comprising, sample liquid analysis meansoperatively connected to said secondary conduit means downstream of saidliquid separating means and operable to analyze the thusly substantiallyseparated first liquid portion.
 9. In apparatus for the introduction ofa first liquid into a stream of a second liquid which is immiscibletherewith and flowing in conduit means, said conduit means comprising aninner conduit means surface which is selectively wettable by said secondliquid to the substantial exclusion of said first liquid, theimprovements comprising, liquid introduction means within said stream ofsaid second liquid in said conduit means for introducing said firstliquid thereinto to form a stream of said first liquid therewithin bydisplacing said second liquid stream substantially to the inner conduitmeans surface for retention thereat by the selective wettability of saidinner conduit means surface by said second liquid to the substantialexclusion of said first liquid, and means operatively connected to saidliquid introduction means for flowing said first liquid therethroughinto said conduit means whereby, said second liquid will substantiallyencapsulate said first liquid stream for continued common flow throughsaid conduit means, said liquid introduction means being fixed relativeto said conduit means.
 10. In apparatus as in claim 9, the improvementsfurther comprising, said liquid introduction means comprising probemeans including a probe means portion which extends within said conduitmeans out of contact with the inner surface of said conduit means, saidprobe means portion comprising outlet means for the flow of said firstliquid out of the same into said second liquid stream.
 11. In apparatusas in claim 10, the improvements further comprising, said means forflowing said first liquid into said conduit means comprising anotherportion of said probe means in communication with said first-mentionedprobe means portion, and extending through the wall of said conduitmeans for the flow of said first liquid into the same.
 12. In apparatusas in claim 10, wherein said inner conduit means surface is hydrophobic,said second liquid is an isolation liquid which is preferentiallyattracted thereof to the substantial exclusion of aqueous sampleliquids, and said first liquid is an aqueous sample liquid which isimmiscible with said isolation liquid, the improvements furthercomprising, sample liquid analysis means operatively connected to saidconduit means downstream of said liquid introduction means and operableto analyze said sample liquid.
 13. In apparatus for the introduction ofa second liquid into a stream of a first liquid which is immiscibletherewith and flowing in conduit means, said conduit means comprising aninner conduit means surface which is selectively wettable by said secondliquid to the substantial exclusion of said first liquid, theimprovements comprising, liquid introduction means in said conduit meansfor introducing said second liquid into said first liquid stream at saidinner conduit means surface to displace said first liquid therefrom andform a stream of said second liquid at said inner conduit means surfacewhcih will be retained thereat by the selective wettability of saidinner conduit means surface by said second liquid to the substantialexclusion of said first liquid, and means operatively connected theretofor flowing said second liquid into said conduit means, whereby saidsecond liquid stream will substantially encapsulate said first liquidstream for continued common flow through said conduit means, said liquidintroduction means comprising a porous conduit means portion of amaterial which is selectively wettable by said second liquid to thesubstantial exclusion of said first liquid, and means operativelyassociated with said porous conduit means portion for flowing saidsecond liquid therethrough into said conduit means.
 14. In apparatus asin claim 13, the improvements further comprising, said porous conduitmeans portion comprising a conduit means insert which is ofsubstantially the same inner diameter as said conduit means.
 15. Inapparatus as in claim 13, the improvements further comprising, saidmeans for flowing said second liquid through said porous conduit meansportion into said conduit means comprise means for pumping the sametherethrough.
 16. In apparatus as in claim 13, the improvements furthercomprising, sleeve means surrounding said porous conduit means portionexternally of the same to create a liquid-tight space therebetween forthe collection of said second liquid prior to the flow thereof throughsaid porous conduit means portion.
 17. In apparatus as in claim 16, theimprovements further comprising, said means to flow said second liquidthrough said porous conduit means portion comprising positivedisplacement pump means operatively connected to said liquid-tight spaceand operable to pump said second liquid thereinto for flow in turnthrough said porous conduit means portion.
 18. In apparatus as in claim13 wherein said inner conduit means surface and said porous conduitmeans portion are hydrophobic, said second liquid is an isolation liquidwhich is preferentially attacted thereto to the substantial exclusion ofaqueous liquids, and said first liquid is an aqueous sample liquid whichis immiscible with said isolation liquid, the improvements furthercomprising, sample liquid analysis means operatively connected to saidconduit means downstream therein of said porous conduit means portionand operable to analyze said first liquid.
 19. In sample liquidsanalysis apparatus including sampler means, sample liquids analysismeans, a source of an isolation liquid which is immiscible with saidsample liquids, and conduit means connecting said sampler means to saidsample liquids analysis means, said sampler means being operable tosupply a stream of successive sample liquids to said conduit means forflow therethrough to said sample liquids analysis means, said conduitmeans comprising an inner surface which is selectively wettable by saidisolation liquid to the substantial exclusion of said sample liquids,the improvements comprising, isolation liquid introduction meansoperatively associated with said conduit means for introducing saidisolation liquid from said isolation liquid source into said sampleliquids stream in said conduit means to form a stream of said isolationliquid at said inner conduit means surface to substantially encapsulatesaid sample liquids stream therein and thereby minimize sample liquidscarryover in said conduit means, and isolation liquid separating meansoperatively associated with said conduit means downstream of saidisolation liquid introduction means for substantially separating saidisolation liquid stream from said sample liquids stream prior to theflow thereof to said sample liquids analysis means, thereby minimizinginterference by said isolation liquid with sample liquids analysis. 20.In apparatus as in claim 19, the improvements further comprising, saidsampler means further comprising means to air segment said sampleliquids stream.
 21. In apparatus as in claim 19, the improvementsfurther comprising, means operatively connected to said isolation liquidseparating means and said isolation liquids source for returning thethusly separated isolation liquid from the former to the latter forre-use in said sample liquids analysis apparatus.
 22. In liquidseparating apparatus for substantially separating at least a portion ofa first liquid from a common liquids stream of first and secondimmiscible liquids flowing in conduit means including an inner conduitmeans surface which is selectively wettable by said second liquid to thesubstantial exclusion of said first liquid, and wherein said firstliquid is flowing primarily within said second liquid substantially outof contact with said conduit means inner surface, the improvementscomprising, liquid separating means in said conduit means forsubstantially separating at least a portion of said first liquid fromsaid common liquids stream attendant the flow thereof past said liquidseparating means in said conduit means, and means operatively connectedto said liquid separating means for flowing the thusly substantiallyseparated first liquid portion out of said conduit means while enablingthe continued flow of the remainder of said common liquids streamthrough said conduit means, said liquid separating means comprisingprobe means including a probe means portion which extends within saidconduit means out of contact with said inner conduit means surface tointercept substantially only the flow of said first liquid in saidconduit means, said probe means portion comprising inlet means forliquid flow thereinto, and said probe means portion comprising an outersurface which is selectively wettable by said second liquid to thesubstantial exclusion of said first liquid.
 23. In liquid separatingapparatus for substantially separating at least a portion of a firstliquid from a common liquids stream of first and second immiscibleliquids flowing in conduit means including an inner conduit meanssurface which is selectively wettable by said second liquid to thesubstantial exclusion of said first liquid, and wherein said firstliquid is flowing primarily within said second liquid substantially outof contact with said conduit means inner surface, the improvementscomprising, liquid separating means in said conduit means forsubstantially separating at least a portion of said first liquid fromsaid common liquids stream attendant the flow thereof past said liquidseparating means in said conduit means, and means operatively connectedthereto for flowing the thusly substantially separated first liquidportion out of said conduit means while enabling the continued flow ofthe remainder of said common liquids stream through said conduit means,said liquid separating means comprising probe means including a probemeans portion which extends within said conduit means out of contactwith said inner conduit means surface to intercept substantially onlythe flow of said first liquid in said conduit means, said probe meansportion comprising inlet means for liquid flow thereinto, said commonliquids stream being air-segmented, and said liquid separating meanscomprising means for separating portions of said air segments in saidcommon liquids stream therefrom for air-segmenting said substantiallyseparated first liquid portion.
 24. In liquid separating apparatus forsubstantially separating at least a portion of a first liquid from acommon liquids stream of first and second immiscible liquids flowing inconduit means including an inner conduit means surface which isselectively wettable by said second liquid to the substantial exclusionof said first liquid, and wherein said first liquid is flowing primarilywithin said second liquid substantially out of contact with said conduitmeans inner surface, the improvements comprising, liquid separatingmeans in said conduit means for substantially separating at least aportion of said first liquid from said common liquids stream attendantthe flow thereof past said liquid separating means in said conduitmeans, and means operatively connected thereto for flowing the thuslysubstantially separated first liquid portion out of said conduit meanswhile enabling the continued flow of the remainder of said commonliquids stream through said conduit means, said liquid separating meanscomprising probe means including a probe means portion which extendswithin said conduit means out of contact with said inner conduit meanssurface to intercept substantially only the flow of said first liquid insaid conduit means, said probe means portion comprising inlet means forliquid flow thereinto, said conduit means being of substantiallyconstant inner diameter coextensive with said conduit means portion. 25.In apparatus for the introduction of a first liquid into a stream of asecond liquid which is immiscible therewith and flowing in conduitmeans, said conduit means comprising an inner conduit means surfacewhich is selectively wettable by said second liquid to the substantialexclusion of said first liquid, the improvements comprising, liquidintroduction means within said stream of said second liquid in saidconduit means for introducing said first liquid thereinto to form astream of said first liquid therewithin by displacing said second liquidstream substantially to the inner conduit means surface for retentionthereat by the selective wettability of said inner conduit means surfaceby said second liquid to the substantial exclusion of said first liquid,means operatively connected to said liquid introduction means forflowing said first liquid therethrough into said conduit means wherebysaid second liquid stream will substantially encapsulate said firstliquid stream for continued common flow through said conduit means, andmeans operatively associated with said first liquid flow means forair-segmenting said first liquid stream.
 26. In apparatus for theintroduction of a first liquid into a stream of a second liquid which isimmiscible therewith and flowing in conduit means, said conduit meanscomprising an inner conduit means surface which is selectively wettableby said second liquid to the substantial exclusion of said first liquid,the improvements comprising, liquid introduction means within saidstream of said second liquid in said conduit means for introducing saidfirst liquid thereinto to form a stream of said first liquid therewithby displacing said second liquid stream substantially to the innerconduit means surface for retention thereat by the selective wettabilityof said inner conduit means surface by said second liquid to thesubstantial exclusion of said first liquid, means operatively connectedto said liquid introduction means for flowing said first liquidtherethrough into said conduit means whereby said second liquid streamwill substantially encapsulate said first liquid stream for continuedcommon flow through said conduit means, said liquid introducing meanscomprising probe means including a probe means portion which extendswithin said conduit means out of contact with the inner surface of saidconduit means, said probe means portion comprising outlet means for theflow of said first liquid out of the same into said second liquidstream, said conduit means and said probe means portion being ofgenerally circular cross-section, with said probe means portion beingdisposed generally concentrically of said conduit means.